Development of a LAMP assay for rapid and sensitive detection and differentiation of Mycobacterium avium subsp. avium and subsp. hominissuis.

Mycobacterium avium causes atypical mycobacterial infection in humans and animals worldwide. M. avium comprises the subspecies avium (MAA), hominissuis (MAH), silvaticum (MAS) and paratuberculosis (MAP). The M. avium complex (MAC), comprising M. avium and M. intracellulare, causes opportunistic infections of humans. M. avium subsp. avium (MAA) mainly causes avian tuberculosis while subsp. hominissuis (MAH) mainly infects pig. Distinguishing between these two subspecies is essential to the effective control of these atypical mycobacterial infections and minimization of the resulting economic loss. For this purpose, we developed a loop-mediated isothermal amplification (LAMP) assay that rapidly and sensitively detects and differentiates MAA and MAH. This MAA-LAMP assay targeting IS901 correctly detected four MAA isolates but did not detect 27 MAH and 19 non-MAA/non-MAH mycobacterial isolates. The MAAH-LAMP assay targeting IS1245 detected four MAA and 27 MAH isolates but not the other 19 mycobacterial isolates. We believe that implementation of this LAMP assay will significantly improve public health and safety. SIGNIFICANCE AND IMPACT OF THE STUDY: Mycobacterium avium, which is pathogenic for humans and animals, represents a continuing threat to public health and safety and to food production. Therefore, improved methods are urgently required to readily and efficiently identify M. avium subspecies. Compared with conventional PCR methods, the LAMP assay herein developed more rapidly detects and better distinguishes between two major M. avium subspecies that cause disease of pig. Importantly, this highly accurate and sensitive LAMP assay detects mycobacterial DNAs using real-time fluorescence or the unaided eye with a colour-change dye, making it ideal for translation to the clinic and slaughterhouse.

[The problem of cardiac surgery with neoplasm].

Neoplasm and cardiac diseases are rarely found at the same time. If the standard treatment for both of these diseases is operation, several problems arise;1 stage or 2 stage operation, which operation should be done 1st etc. Especially, if cardiac operation needs cardiopulmonary bypass, neoplasm may grow faster. In such cases, we must decide operative procedure carefully.

[One-stage coronary artery bypass grafting and revascularization in a patient with ischemic heart disease and arteriosclerosis obliterans].

With the overall increase in ischemic heart disease (IHD), cases combining arteriosclerosis obliterans (ASO) of the lower extremities and IHD are on the rise. Therefore, it is necessary to consider methods and timing of operation. These cases have occasionally large collateral pathways from the internal thoracic artery (ITA) to the femoral artery via the epigastric artery. To prevent irreversible ischemia of the lower limbs after harvesting of ITA, we planned to perform a one-stage operation for ASO-combined IHD. Revascularization of the lower extremities was performed 1st. Subsequently coronary artery bypass grafting with ITA was completed. The operative courses were uneventful.

[Chondrosarcoma of the rib with pleural dissemination].

A 52-year-old woman was admitted to the hospital because of abnormal shadow of the left lung. Chest computed tomography (CT) showed a 10.0 x 6.6 cm mass arising from the left chest wall. The diagnosis of grade II chondrosarcoma was made with CT guided percutaneous needle biopsy. At operation, the irregularly-shaped tumor was arising from the 7th rib. Chest wall resection and partial resection of the left lower lobe including the tumor was performed. Multiple miliary disseminations were noted on the pleura and complete removal of these miliary lesions was impossible. Adjuvant chemotherapy using OK-432, carboplatin and etoposide was tried as treatment for pleural dissemination. No signs of disease progression have been seen as of 12 months after the operation. Radical excision of chondrosarcoma of the rib including all pleural dissemination is difficult. Multidisciplinary approach including resection of a primary tumor has potential to improve prognosis in some cell types. And long term follow-up is necessary to determine effectiveness of adjuvant chemotherapy in this case.

[Intentional limited resection for lung adenocarcinoma of 2 cm in size or less according to intraoperative frozen section and high-resolution computed tomography].

One hundred twenty-six cases of resected lung adenocarcinoma of 2 cm in size or less were studied about intra and postoperative Noguchi's classification, high-resolution computed tomography (HRCT) findings and lymph node metastases in cases of lobectomy. Intentional limited resection for lung adenocarcinoma of 2 cm in size or less was recommended for selected patients with tumors of type A of Noguchi's classification diagnosed by intraoperative frozen diagnosis or tumors of intraoperative type B of 0.5 in ground-glass opacity (GGO) diameter ratio or more of HRCT. These patients are almost equal to patients of 0.5 in GGO diameter ratio or more with any type of Noguchi's classification. This GGO diameter ratio of 0.5 is easily available as a criteria of intentional limited resection for lung adenocarcinoma of 2 cm in size or less.

[Right atrial blood cyst; report of a case].

A 53-year-old female was admitted to our hospital for examination of an abnormal shadow on chest X-ray. Computed tomography (CT) revealed a large mass of 3 cm in diameter in the right atrium. The mass arose from the right atrial septum and was removed with the interatrial septum. Histological specimen showed a blood cyst contained serous effusion.

Chemiluminescence of whole blood. II. Application to clinical examination of phagocytic functions of whole blood from various types of disease.

Measurements of whole blood chemiluminescence (CL) were carried out by using 0.1 ml of blood to evaluate phagocytic functions from various kinds of pediatric patients. Patients with definite bacterial infections showed an elevated background CL and high peak CL per 1000 granulocytes after stimulation with zymosan. These results suggest that granulocytes of these patients had been preactivated in vivo, their phagocytic and/or metabolic activity being enhanced. Contrary to these results, blood from the patients with viral infections showed almost no abnormalities. Patients with chronic granulomatous disease showed no significant CL by this method similar to conventional ones. This method was also shown to be a useful technique for monitoring the phagocytic functions of blood after granulocyte transfusion.